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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 郭錦樺(Ching-Hua Kuo) | |
dc.contributor.author | Wei-Chieh Wang | en |
dc.contributor.author | 王韋傑 | zh_TW |
dc.date.accessioned | 2023-03-19T22:28:11Z | - |
dc.date.copyright | 2022-10-05 | |
dc.date.issued | 2022 | |
dc.date.submitted | 2022-08-30 | |
dc.identifier.citation | 1. Han, X. and H. Ye, Overview of Lipidomic Analysis of Triglyceride Molecular Species in Biological Lipid Extracts. J Agric Food Chem, 2021. 69(32): p. 8895-8909. 2. Luna-Castillo, K.P., et al., The Effect of Dietary Interventions on Hypertriglyceridemia: From Public Health to Molecular Nutrition Evidence. Nutrients, 2022. 14(5). 3. Zechner, R., et al., Adipose triglyceride lipase and the lipolytic catabolism of cellular fat stores. J Lipid Res, 2009. 50(1): p. 3-21. 4. Alves-Bezerra, M. and D.E. Cohen, Triglyceride Metabolism in the Liver. Compr Physiol, 2017. 8(1): p. 1-8. 5. Wiesner, P. and K.E. Watson, Triglycerides: A reappraisal. Trends Cardiovasc Med, 2017. 27(6): p. 428-432. 6. Yang, K. and X. Han, Lipidomics: Techniques, Applications, and Outcomes Related to Biomedical Sciences. Trends Biochem Sci, 2016. 41(11): p. 954-969. 7. Manne, V., P. Handa, and K.V. Kowdley, Pathophysiology of Nonalcoholic Fatty Liver Disease/Nonalcoholic Steatohepatitis. Clin Liver Dis, 2018. 22(1): p. 23-37. 8. Liebisch, G., et al., Update on LIPID MAPS classification, nomenclature, and shorthand notation for MS-derived lipid structures. J Lipid Res, 2020. 61(12): p. 1539-1555. 9. Bayly, G.R., CHAPTER 37 - Lipids and disorders of lipoprotein metabolism, in Clinical Biochemistry: Metabolic and Clinical Aspects (Third Edition), W.J. Marshall, et al., Editors. 2014, Churchill Livingstone. p. 702-736. 10. Bates, P.D. and J. Browse, The significance of different diacylgycerol synthesis pathways on plant oil composition and bioengineering. Front Plant Sci, 2012. 3: p. 147. 11. Yen, C.L., et al., Thematic review series: glycerolipids. DGAT enzymes and triacylglycerol biosynthesis. J Lipid Res, 2008. 49(11): p. 2283-301. 12. Jensen-Urstad, A.P. and C.F. Semenkovich, Fatty acid synthase and liver triglyceride metabolism: housekeeper or messenger? Biochim Biophys Acta, 2012. 1821(5): p. 747-53. 13. Tada, H., A. Nohara, and M.A. Kawashiri, Serum Triglycerides and Atherosclerotic Cardiovascular Disease: Insights from Clinical and Genetic Studies. Nutrients, 2018. 10(11). 14. Hidalgo, N.J., et al., Elevated Serum Triglyceride Levels in Acute Pancreatitis: A Parameter to be Measured and Considered Early. World J Surg, 2022. 15. Georgiopoulou, V.V., et al., Metabolic abnormalities in adults with cystic fibrosis. Respirology, 2010. 15(5): p. 823-9. 16. Low, S., et al., The role of triglyceride glucose index in development of Type 2 diabetes mellitus. Diabetes Res Clin Pract, 2018. 143: p. 43-49. 17. Yang, A.L. and J. McNabb-Baltar, Hypertriglyceridemia and acute pancreatitis. Pancreatology, 2020. 20(5): p. 795-800. 18. Ishimo, M.C., et al., Hypertriglyceridemia is associated with insulin levels in adult cystic fibrosis patients. J Cyst Fibros, 2013. 12(3): p. 271-6. 19. Xu, S.L., et al., Research advances based on mass spectrometry for profiling of triacylglycerols in oils and fats and their applications. Electrophoresis, 2018. 39(13): p. 1558-1568. 20. Beckles, D.M. and U. Roessner, Plant metabolomics, in Plant Biotechnology and Agriculture. 2012. p. 67-81. 21. Liu, T., et al., LC-MS-based lipid profile in colorectal cancer patients: TAGs are the main disturbed lipid markers of colorectal cancer progression. Anal Bioanal Chem, 2019. 411(20): p. 5079-5088. 22. Nambiar, S., et al., There is detectable variation in the lipidomic profile between stable and progressive patients with idiopathic pulmonary fibrosis (IPF). Respir Res, 2021. 22(1): p. 105. 23. Camera, E., et al., Comprehensive analysis of the major lipid classes in sebum by rapid resolution high-performance liquid chromatography and electrospray mass spectrometry. J Lipid Res, 2010. 51(11): p. 3377-88. 24. Ackerman, D., et al., Triglycerides Promote Lipid Homeostasis during Hypoxic Stress by Balancing Fatty Acid Saturation. Cell Rep, 2018. 24(10): p. 2596-2605 e5. 25. Tose, L.V., et al., Coupling Stable Isotope Labeling and Liquid Chromatography-Trapped Ion Mobility Spectrometry-Time-of-Flight-Tandem Mass Spectrometry for De Novo Mosquito Ovarian Lipid Studies. Anal Chem, 2022. 94(16): p. 6139-6145. 26. Triebl, A., et al., Lipidomics: Prospects from a technological perspective. Biochim Biophys Acta Mol Cell Biol Lipids, 2017. 1862(8): p. 740-746. 27. Bird, S.S., et al., Serum lipidomics profiling using LC-MS and high-energy collisional dissociation fragmentation: focus on triglyceride detection and characterization. Anal Chem, 2011. 83(17): p. 6648-57. 28. Chung, K.P., et al., Multi-kinase framework promotes proliferation and invasion of lung adenocarcinoma through activation of dynamin-related protein 1. Mol Oncol, 2021. 15(2): p. 560-578. 29. Folch, J., M. Lees, and G.H. Sloane Stanley, A simple method for the isolation and purification of total lipids from animal tissues. J biol Chem, 1957. 226(1): p. 497-509. 30. Benjamini, Y. and Y. Hochberg, Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing. Journal of the Royal Statistical Society: Series B (Methodological), 1995. 57(1): p. 289-300. 31. Pang, Z., et al., MetaboAnalyst 5.0: narrowing the gap between raw spectra and functional insights. Nucleic Acids Res, 2021. 49(W1): p. W388-W396. 32. Aristizabal-Henao, J.J., et al., Nontargeted lipidomics of novel human plasma reference materials: hypertriglyceridemic, diabetic, and African-American. Anal Bioanal Chem, 2020. 412(27): p. 7373-7380. 33. Tkachev, A., et al., Shorter Chain Triglycerides Are Negatively Associated with Symptom Improvement in Schizophrenia. Biomolecules, 2021. 11(5). 34. Le, H., M. Jerums, and C.T. Goudar, Characterization of intrinsic variability in time-series metabolomic data of cultured mammalian cells. Biotechnol Bioeng, 2015. 112(11): p. 2276-83. 35. Crews, B., et al., Variability Analysis of Human Plasma and Cerebral Spinal Fluid Reveals Statistical Significance of Changes in Mass Spectrometry-Based Metabolomics Data. Analytical Chemistry, 2009. 81(20): p. 8538-8544. 36. Wu, Y. and L. Li, Sample normalization methods in quantitative metabolomics. J Chromatogr A, 2016. 1430: p. 80-95. 37. Ryan, D., et al., Recent and potential developments in the analysis of urine: a review. Anal Chim Acta, 2011. 684(1-2): p. 8-20. 38. Ferchaud-Roucher, V., et al., Plasma Lipidome Analysis by Liquid Chromatography-High Resolution Mass Spectrometry and Ion Mobility of Hypertriglyceridemic Patients on Extended-Release Nicotinic Acid: a Pilot Study. Cardiovasc Drugs Ther, 2017. 31(3): p. 269-279. 39. Ferchaud-Roucher, V., et al., Omega 3 Improves Both apoB100-containing Lipoprotein Turnover and their Sphingolipid Profile in Hypertriglyceridemia. J Clin Endocrinol Metab, 2020. 105(10). 40. Croyal, M., et al., Effects of Extended-Release Nicotinic Acid on Apolipoprotein (a) Kinetics in Hypertriglyceridemic Patients. Arterioscler Thromb Vasc Biol, 2015. 35(9): p. 2042-7. 41. Chung, K.P., et al., Mitofusins regulate lipid metabolism to mediate the development of lung fibrosis. Nat Commun, 2019. 10(1): p. 3390. 42. Whitsett, J.A., S.E. Wert, and T.E. Weaver, Alveolar surfactant homeostasis and the pathogenesis of pulmonary disease. Annu Rev Med, 2010. 61: p. 105-19. 43. Lass, A., et al., Lipolysis - a highly regulated multi-enzyme complex mediates the catabolism of cellular fat stores. Prog Lipid Res, 2011. 50(1): p. 14-27. 44. Simha, V., Management of hypertriglyceridemia. BMJ, 2020. 371: p. m3109. 45. Miri, R., et al., Alterations in oxidative stress biomarkers associated with mild hyperlipidemia and smoking. Food Chem Toxicol, 2012. 50(3-4): p. 920-6. 46. Zhang, A., et al., A Study on the Factors Influencing Triglyceride Levels among Adults in Northeast China. Sci Rep, 2018. 8(1): p. 6388. 47. Ruiz-Garcia, A., et al., Prevalence of hypertriglyceridemia in adults and related cardiometabolic factors. SIMETAP-HTG study. Clin Investig Arterioscler, 2020. 32(6): p. 242-255. 48. Valdivielso, P., A. Ramirez-Bueno, and N. Ewald, Current knowledge of hypertriglyceridemic pancreatitis. Eur J Intern Med, 2014. 25(8): p. 689-94. 49. Packard, C.J., J. Boren, and M.R. Taskinen, Causes and Consequences of Hypertriglyceridemia. Front Endocrinol (Lausanne), 2020. 11: p. 252. 50. Reiner, Z., Hypertriglyceridaemia and risk of coronary artery disease. Nat Rev Cardiol, 2017. 14(7): p. 401-411. 51. Brahm, A. and R.A. Hegele, Hypertriglyceridemia. Nutrients, 2013. 5(3): p. 981-1001. 52. Laufs, U., et al., Clinical review on triglycerides. Eur Heart J, 2020. 41(1): p. 99-109c. 53. Sunil, B. and A.P. Ashraf, Childhood Hypertriglyceridemia: Is It Time for a New Approach? Curr Atheroscler Rep, 2022. 54. Nordestgaard, B.G. and A. Varbo, Triglycerides and cardiovascular disease. The Lancet, 2014. 384(9943): p. 626-635. 55. Pejic, R.N. and D.T. Lee, Hypertriglyceridemia. The Journal of the American Board of Family Medicine, 2006. 19(3): p. 310-316. 56. Yuan, G., K.Z. Al-Shali, and R.A. Hegele, Hypertriglyceridemia: its etiology, effects and treatment. CMAJ, 2007. 176(8): p. 1113-20. 57. Peterson, B.E., et al., Reduction in Revascularization With Icosapent Ethyl: Insights From REDUCE-IT Revascularization Analyses. Circulation, 2021. 143(1): p. 33-44. 58. Mason, R.P., P. Libby, and D.L. Bhatt, Emerging Mechanisms of Cardiovascular Protection for the Omega-3 Fatty Acid Eicosapentaenoic Acid. Arterioscler Thromb Vasc Biol, 2020. 40(5): p. 1135-1147. 59. Giammanco, A., et al., The pathophysiology of intestinal lipoprotein production. Front Physiol, 2015. 6: p. 61. 60. Tiwari, S. and S.A. Siddiqi, Intracellular Trafficking and Secretion of VLDL. Arteriosclerosis, Thrombosis, and Vascular Biology, 2012. 32(5): p. 1079-1086. 61. Xu, Y., et al., Targeting ApoC3 Paradoxically Aggravates Atherosclerosis in Hamsters With Severe Refractory Hypercholesterolemia. Front Cardiovasc Med, 2022. 9: p. 840358. 62. Gonzales, J.C., et al., Apolipoproteins E and AV mediate lipoprotein clearance by hepatic proteoglycans. J Clin Invest, 2013. 123(6): p. 2742-51. 63. Wilson, D.E., et al., Phenotypic expression of heterozygous lipoprotein lipase deficiency in the extended pedigree of a proband homozygous for a missense mutation. J Clin Invest, 1990. 86(3): p. 735-50. 64. Peterfy, M., et al., Mutations in LMF1 cause combined lipase deficiency and severe hypertriglyceridemia. Nat Genet, 2007. 39(12): p. 1483-7. 65. Beigneux, A.P., et al., Glycosylphosphatidylinositol-anchored high-density lipoprotein-binding protein 1 plays a critical role in the lipolytic processing of chylomicrons. Cell Metab, 2007. 5(4): p. 279-91. 66. Mahley, R.W. and S.C. Rall, Jr., Apolipoprotein E: far more than a lipid transport protein. Annu Rev Genomics Hum Genet, 2000. 1: p. 507-37. 67. Matsunaga, A., et al., Variants of Lipid-Related Genes in Adult Japanese Patients with Severe Hypertriglyceridemia. J Atheroscler Thromb, 2020. 27(12): p. 1264-1277. 68. Hegele, R.A., et al., The polygenic nature of hypertriglyceridaemia: implications for definition, diagnosis, and management. The Lancet Diabetes & Endocrinology, 2014. 2(8): p. 655-666. 69. Lee, S.Y., et al., Altered plasma lysophosphatidylcholines and amides in non-obese and non-diabetic subjects with borderline-to-moderate hypertriglyceridemia: a case-control study. PLoS One, 2015. 10(4): p. e0123306. 70. Li, B., et al., Untargeted fecal metabolomics revealed biochemical mechanisms of the blood lipid-lowering effect of koumiss treatment in patients with hyperlipidemia. Journal of Functional Foods, 2021. 78. 71. Rai, S. and S. Bhatnagar, Novel Lipidomic Biomarkers in Hyperlipidemia and Cardiovascular Diseases: An Integrative Biology Analysis. OMICS, 2017. 21(3): p. 132-142. 72. Catapano, A.L., et al., 2016 ESC/EAS Guidelines for the Management of Dyslipidaemias. Eur Heart J, 2016. 37(39): p. 2999-3058. 73. Duan, D.M., et al., Clinical manifestations and genetic characteristics in the Taiwan thoracic aortic aneurysm and dissection cohort - a prospective cohort study. J Formos Med Assoc, 2022. 121(6): p. 1093-1101. 74. Hsiung, Y.C., et al., Identification of a novel LDLR disease-causing variant using capture-based next-generation sequencing screening of familial hypercholesterolemia patients in Taiwan. Atherosclerosis, 2018. 277: p. 440-447. 75. Li, H. and R. Durbin, Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics, 2009. 25(14): p. 1754-60. 76. Karczewski, K.J., et al., The mutational constraint spectrum quantified from variation in 141,456 humans. Nature, 2020. 581(7809): p. 434-443. 77. Richards, S., et al., Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Med, 2015. 17(5): p. 405-24. 78. Scherer, J., et al., Issues in hypertriglyceridemic pancreatitis: an update. J Clin Gastroenterol, 2014. 48(3): p. 195-203. 79. Charlesworth, A., A. Steger, and M.A. Crook, Acute pancreatitis associated with severe hypertriglyceridaemia; A retrospective cohort study. Int J Surg, 2015. 23(Pt A): p. 23-7. 80. Pedersen, S.B., A. Langsted, and B.G. Nordestgaard, Nonfasting Mild-to-Moderate Hypertriglyceridemia and Risk of Acute Pancreatitis. JAMA Intern Med, 2016. 176(12): p. 1834-1842. 81. Mariani, A. and P.A. Testoni, Is acute recurrent pancreatitis a chronic disease? World J Gastroenterol, 2008. 14(7): p. 995-8. 82. Ahmed Ali, U., et al., Risk of Recurrent Pancreatitis and Progression to Chronic Pancreatitis After a First Episode of Acute Pancreatitis. Clin Gastroenterol Hepatol, 2016. 14(5): p. 738-46. 83. Bachorik, P.S. and J.W. Ross, National Cholesterol Education Program recommendations for measurement of low-density lipoprotein cholesterol: executive summary. The National Cholesterol Education Program Working Group on Lipoprotein Measurement. Clin Chem, 1995. 41(10): p. 1414-20. 84. Shimizu, M., et al., Very long chain fatty acids are an important marker of nutritional status in patients with anorexia nervosa: a case control study. Biopsychosoc Med, 2020. 14: p. 14. 85. Kihara, A., Very long-chain fatty acids: elongation, physiology and related disorders. J Biochem, 2012. 152(5): p. 387-95. 86. Jacobs, S., et al., Associations of Erythrocyte Fatty Acids in the De Novo Lipogenesis Pathway with Proxies of Liver Fat Accumulation in the EPIC-Potsdam Study. PLoS One, 2015. 10(5): p. e0127368. 87. Song, Z., A.M. Xiaoli, and F. Yang, Regulation and Metabolic Significance of De Novo Lipogenesis in Adipose Tissues. Nutrients, 2018. 10(10). 88. Wu, J.H., et al., Fatty acids in the de novo lipogenesis pathway and risk of coronary heart disease: the Cardiovascular Health Study. Am J Clin Nutr, 2011. 94(2): p. 431-8. 89. M K Hellerstein, a. J-M Schwarz, and R.A. Neese, Regulation of Hepatic De Novo Lipogenesis in Humans. Annual Review of Nutrition, 1996. 16(1): p. 523-557. 90. Imamura, F., et al., Fatty acids in the de novo lipogenesis pathway and incidence of type 2 diabetes: A pooled analysis of prospective cohort studies. PLoS Med, 2020. 17(6): p. e1003102. 91. Roumans, K.H.M., et al., Hepatic saturated fatty acid fraction is associated with de novo lipogenesis and hepatic insulin resistance. Nat Commun, 2020. 11(1): p. 1891. 92. Fritsche, K.L., The science of fatty acids and inflammation. Adv Nutr, 2015. 6(3): p. 293S-301S. 93. Calder, P.C., Long-chain fatty acids and inflammation. Proc Nutr Soc, 2012. 71(2): p. 284-9. 94. Gupta, S., et al., Saturated long-chain fatty acids activate inflammatory signaling in astrocytes. J Neurochem, 2012. 120(6): p. 1060-71. 95. Listenberger, L.L., et al., Triglyceride accumulation protects against fatty acid-induced lipotoxicity. Proceedings of the National Academy of Sciences, 2003. 100(6): p. 3077-3082. 96. Alamri, H., et al., Mapping the triglyceride distribution in NAFLD human liver by MALDI imaging mass spectrometry reveals molecular differences in micro and macro steatosis. Analytical and Bioanalytical Chemistry, 2018. 411(4): p. 885-894. 97. Saini, R.K. and Y.S. Keum, Omega-3 and omega-6 polyunsaturated fatty acids: Dietary sources, metabolism, and significance - A review. Life Sci, 2018. 203: p. 255-267. 98. Skulas-Ray, A.C., et al., Omega-3 Fatty Acids for the Management of Hypertriglyceridemia: A Science Advisory From the American Heart Association. Circulation, 2019. 140(12): p. e673-e691. 99. Tremblay, A.J., et al., Associations between the fatty acid content of triglyceride, visceral adipose tissue accumulation, and components of the insulin resistance syndrome. Metabolism, 2004. 53(3): p. 310-7. 100. Kotronen, A., et al., Serum saturated fatty acids containing triacylglycerols are better markers of insulin resistance than total serum triacylglycerol concentrations. Diabetologia, 2009. 52(4): p. 684-90. 101. Yang, K. and X. Han, Accurate quantification of lipid species by electrospray ionization mass spectrometry - Meet a key challenge in lipidomics. Metabolites, 2011. 1(1): p. 21-40. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/84836 | - |
dc.description.abstract | 三酸甘油脂 (Triglyceride, TG) 做為人體內脂肪運輸與能量儲存的主要分子。TG大於 150 mg/dL是目前診斷高三酸甘油脂血症 (Hypertriglyceridemia, HTG) 的標準,且被認為有較高風險罹患急性胰臟炎 (Acute pancreatitis, AP)、二型糖尿病 (Type 2 diabetes mellitus, T2DM) 與心血管疾病 (Cardiovascular disease, CVD)。在臨床上,三酸甘油脂僅被用以當作診斷標準,且因支鏈脂肪酸 (Fatty acid, FA) 所產生的複雜結構導致分析困難而使不同支鏈組成所造成的潛在影響尚未明瞭。本研究致力於開發以質譜為主要分析工具之偽標的分析平台以促進三酸甘油脂之研究。 於第一章節中,我們開發使用液相層析串聯質譜系統的三酸甘油脂分析流程與平台。我們最佳化樣品配置流程以完整萃取樣品中的三酸甘油脂並降低基質效應 (Metric effect);同時最佳化液相層析參數以獲得良好分離效果。全圖譜掃描 (Full scan mode) 被用以取得各式樣品間不同的三酸甘油脂種類列表,並藉由子離子掃描 (Product ion scan, PIS) 以獲得個別分子中的支鏈組成。這些子母離子可組合成多反應監測 (Multiple reaction monitoring, MRM) 離子對 (Transition)。層析結果顯示甲醇 (Methanol, MeOH)與異丙醇 (Isopropyl alcohol, IPA)的混合溶液可完整溶解樣品且Kinetex® 碳18管柱(2.6µm; 50 x 2.1mm) 提供良好層析圖譜。在現有的層析條件下,所有三酸甘油脂可依其碳當量 (Equivalent carbon number, ECN) 序列析出。本實驗挑選兩種樣品進行平台測試,分別為剪除脂肪酸合成蛋白 (Fatty acid synthase, FASN) 基因之鼠肺泡上皮細胞2型 (Murine type 2 alveolar epithelial cells, AEC2) 細胞株-MLE-12細胞與高三酸甘油脂血症患者血漿,上述兩者分別代表不同種類之樣品。在兩種不同的樣品底下我們偵測到不同的三酸甘油脂種類列表與不同的支鏈組成,發現對於三酸甘油脂組成分析的研究,針對樣品先進行TG種類篩選的重要性。 第二章節中,我們分析頑固性與非頑固性高三酸甘油脂血症患者血漿,並觀察到兩者血漿呈現特殊差異趨勢。其中我們將嚴重三酸甘油脂血症患者依其服用降血脂藥後之反應分成頑固性與非頑固性患者 (頑固性患者服藥後體內三酸甘油脂濃度持續高於150 mg/dL)。四種與脂蛋白酶和脂蛋白代謝相關基因:LPL、LMF1、ApoA1、ApoA5,亦被收錄以評測基因對頑固性症狀之影響。 140份血漿樣品中,80份來自頑固性患者、20份來自非頑固性患者、40份來自健康對照組。其血漿中三酸甘油脂含量分別為:784.4 ± 818.2 mg/dL、116.8 ± 28.1 mg/dL、和 84.9 ± 38.5 mg/dL。偏最小平方判別分析 (Partial least squares-discriminant analysis, PLS-DA) 顯示頑固性高三酸甘油脂血症患者有較低比例的高不飽和度三酸甘油脂。在經過用藥紀錄的校正後,17 個三酸甘油脂中的支鏈在三酸甘油脂血症患者血漿中有顯著性的較低比例,且其中有多個omega-6 類型的脂肪酸。再者,若將TG 19:1_34:2 (OR: 0.971; 95% CI: 0.94-0.99) 和TG 18:2_38:5 (OR: 0.998; 95% CI: 0.99-1.00) 兩個三酸甘油脂佐以用藥紀錄可有效區分高三酸甘油脂血症中的頑固性症狀患者,其曲線下面積 (Area under the receiver operating characteristic, AUROC) 可從0.826上升到0.944。 本研究開發一個高效的偽標的分析平台並藉由兩種樣品模型展示平台之應用性。我們同時發現頑固性高三酸甘油脂血症之患者體內有較低比例含多不飽和支鏈的三酸甘油脂。這個發現有助於在臨床鑑定潛在的頑固性症狀患者。然而,僅作為一個探索性實驗 (Pilot test),本現象仍須更多的關注與研究以鑑定頑固性症狀背後不同三酸甘油脂組成之影響。 | zh_TW |
dc.description.abstract | Triglyceride (TG) is essential in transporting dietary fat and storing energy. Hypertriglyceridemia (HTG) is a metabolic disease characterized by fasting TG levels above 150 mg/dL, and it is believed to be related to various diseases such as acute pancreatitis (AP), type 2 diabetes mellitus (T2DM), and cardiovascular diseases (CVD). Conventionally, TGs are only regarded as a whole in clinical diagnosis; the underlying role of different TG compositions remains unclear. Nonetheless, the structural complexity of three fatty acids (FA) chains makes TGs analysis challenging. The study aims to develop an LC-MS based pseudo-target TG analytical platform to facilitate TG analysis. In the first chapter, the study aimed to develop a TG analytical platform using LC-MS systems. The sample preparation protocol was optimized to thoroughly extract the sample and alleviate the metric effect, and LC parameters such as analytical column, gradient, and flow rate were optimized to provide good separation. Targeted TG species lists were acquired by full scan mode, while the product ion scan (PIS) was used to identify fatty acyl chains. The results were transformed into multiple reaction monitoring (MRM) transitions. The LC chromatogram suggested that the mixed solvent of methanol (MeOH) and isopropyl alcohol (IPA) could completely dissolve the extracted samples, and Kinetex® C18 Column (2.6µm; 50 x 2.1mm) provided better separation efficacy. Moreover, the optimized gradient profile could provide favorable linear correlations between equivalent carbon number (ECN) and retention time. Two demonstration samples, murine type 2 alveolar epithelial cells (AEC2) line, MLE-12, with Fasn gene deletion and hypertriglyceridemia (HTG) plasma, were chosen to display the capability of the platform. The finding of different TG species lists strengthened the importance of TG target list acquisition. The second chapter investigated TG compositions of severe HTG patients with and without refractory. The refractory was diagnosed as the patients had TG levels above 150 mg/dL after medical treatment. Four coding regions related to lipoprotein lipase (LPL) and apolipoprotein functionality, including LPL, LMF1, ApoA1, and ApoA5 genes, were sequenced in HTG patients to evaluate the genetic effects. Of 140 enrolled subjects, there were 80 refractory HTG (rHTG) patients, 20 non-refractory HTG (nrHTG) patients, and 40 normal controls. The TG levels of three groups are 784.4 ± 818.2 mg/dL, 116.8 ± 28.1 mg/dL, and 84.9 ± 38.5 mg/dL, respectively. The PLS-DA plot revealed that the rHTG patients had a lower proportion of TG with higher unsaturated degrees. In addition, 17 TG compositions showed a significantly lower proportion in rHTG after adjusting drug use, and the omega-6 FAs were commonly listed within these TG compositions. After incorporating TG biomarkers of TG 19:1_34:2 (OR: 0.971; 95% CI: 0.94-0.99) and TG 18:2_38:5 (OR: 0.998; 95% CI: 0.99-1.00) with drug use to identify patients with rHTG, the area under the receiver operating characteristic (AUROC) increased from 0.826 to 0.944. This study developed an effective TG analytical platform with the demonstration of two model samples. We also presented that rHTG patients processed a lower proportion of TG compositions with poly-unsaturated FA. The results may help differentiate the potential refractory severe HTG patients in clinical practice. Nevertheless, as a pilot study, more studies are needed to elucidate the mechanism of different TG compositions with refractory. | en |
dc.description.provenance | Made available in DSpace on 2023-03-19T22:28:11Z (GMT). No. of bitstreams: 1 U0001-1008202211125300.pdf: 3109529 bytes, checksum: d6d880a4611f8eb68ac1f9379bce5eeb (MD5) Previous issue date: 2022 | en |
dc.description.tableofcontents | 誌謝 I 摘要 III Abstract V Content VII Figure content IX Table content XI Chapter 1 Using a pseudo-targeted profiling strategy for studying triglyceride regulation in biologics 1. Introduction 2 2. Materials and methods 6 2.1 Chemical and instrument 6 2.2 Cell culture 6 2.3 Immunoblot 7 2.4 Sample preparation and pre-normalization 8 2.5 LC/MS and analytical workflow 8 2.6 Analytical procedure 9 2.7 Data processing and statistical analysis 9 3. Result 11 3.1 Method optimization 11 3.2 TG analytical workflow establishment 13 3.3 Application 14 4. Discussion 16 5. Conclusion 19 6. Figure 21 7. Table 34 Chapter 2 Plasma triglyceride profiling reveals a lower proportion of unsaturated triglyceride in patients with refractory severe hypertriglyceridemia 1. Introduction 45 2. Materials and methods 48 2.1 Disclaimer 48 2.2 Patient recruitment 48 2.3 Exclusion criteria 49 2.4 Research subject and study design 49 2.5 HTG-related gene detection 49 2.5.1 Hybridization capture-based target enrichment – probes 50 2.5.2 Genomic DNA (GDNA) Extraction 50 2.5.3 Sequencing 50 2.5.4 Read mapping and variant calling 51 2.5.5 Filtering 51 2.5.6 Variant classification 51 2.6 TG compositions profiling in HTG patients by LC-MS 52 2.6.1 Sample preparation 52 2.6.2 Analytical workflow and LC-MS parameter 52 2.6.3 LC-MS data processing and statistical analysis 53 3. Result 54 3.1 The comparison of HTG and NTG 55 3.2 The comparison of rsHTG and nrHTG 55 3.3 The comparison of rsHTG, rHTG, nrHTG and NTG 56 3.4 Biomarker discovery 57 4. Discussion 57 5. Conclusion 63 6. Figure 64 7. Table 68 8. Reference 71 | |
dc.language.iso | en | |
dc.title | 以液相層析四極柱質譜儀開發三酸甘油脂之分析方法並分析2型肺泡上皮細胞及高三酸甘油脂血症患者血漿 | zh_TW |
dc.title | Development of Triglyceride Profiling Method and the Application on Studying Alveolar Epithelial Type 2 Cells and Hypertriglyceridemia Plasma by LC-MS/MS | en |
dc.type | Thesis | |
dc.date.schoolyear | 110-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 鐘桂彬(Kuei-Pin Chung) | |
dc.contributor.oralexamcommittee | 陳沛隆(Pei-Lung Chen),林靖愉(Ching-Yu Lin) | |
dc.subject.keyword | 偽標的分析,液相層析串聯質譜儀,鼠肺泡上皮細胞二型,脂肪酸合成蛋白,三酸甘油脂,高三酸甘油脂血症, | zh_TW |
dc.subject.keyword | Pseudo-target analysis,Liquid chromatography-mass spectrometry (LC-MS),Murine type 2 alveolar epithelial cells (AEC2),Fatty acid synthase (FASN),Triglyceride (TG),Hypertriglyceridemia (HTG), | en |
dc.relation.page | 78 | |
dc.identifier.doi | 10.6342/NTU202202244 | |
dc.rights.note | 同意授權(限校園內公開) | |
dc.date.accepted | 2022-08-30 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 藥學研究所 | zh_TW |
dc.date.embargo-lift | 2027-08-26 | - |
顯示於系所單位: | 藥學系 |
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